[0002]The identification of a therapeutic compound effective for the
prophylaxis and/or treatment of a disease can be based on the activity of
the compound in a biological assay. A biological assay that mimics a
disease causative mechanism can be used to test the therapeutic activity
of a candidate peptide.

[0003]The causative mechanism of many diseases is the over activity of a
biological pathway. A peptide that can reduce the activity of the
biological pathway can be effective in the prophylaxis and/or treatment
of the disease caused by the over activity of the biological pathway.
Similarly the causative mechanism of many diseases is the over production
of a biological molecule. A peptide that can reduce the production of the
biological molecule or block the activity of the over produced biological
molecule can be effective in the prophylaxis and/or treatment of the
disease caused by the over production of the biological molecule.

[0004]Conversely, the causative mechanism of many diseases is the under
activity of a biological pathway. A peptide that can increase the
activity of the biological pathway can be effective in the prophylaxis
and/or treatment of the disease caused by the under activity of the
biological pathway. Also similarly the causative mechanism of many
diseases is the under production of a biological molecule. A peptide that
can increase the production of the biological, molecule or mimic the
biological activity of the under produced biological molecule can be
effective in the prophylaxis and/or treatment of the disease caused by
the under production of the biological molecule.

[0006]The object of the present invention is solved by the teaching of the
independent claims. Further advantageous features, aspects and details of
the invention are evident from the dependent claims, the description, and
the examples of the present application.

[0009]The peptide of the present invention was tested using the assays
described in Examples 1-7, 9-18 for their effect as active therapeutic
agents in the prophylaxis and/or treatment of cancer, proliferative
diseases, tumors and their metastases.

Infectious Disease

[0010]The immune system in higher vertebrates represents the first line of
defense against various antigens that can enter the vertebrate body,
including microorganisms such as bacteria, fungi and viruses that are the
causative agents of a variety of diseases.

[0011]Despite large immunization programs, viral infections, such as
influenza virus, human immunodeficiency virus ("HIV"), herpes simplex
virus ("HSV", type 1 or 2), human papilloma virus ("HPV", type 16 or 18),
human cytomegalovirus ("HCMV") or human hepatitis B or C virus ("HBV",
Type B; "HCV", type C) infections, remain a serious source of morbidity
and mortality throughout the world and a significant cause of illness and
death among people with immune-deficiency associated with aging or
different clinical conditions. Although antiviral chemotherapy with
compounds such as amantadine and rimantadine have been shown to reduce
the duration of symptoms of clinical infections (i.e., influenza
infection), major side effects and the emergence of drug-resistant
variants have been described. New classes of antiviral agents designed to
target particular viral proteins such as influenza neuraminidase are
being developed. However, the ability of viruses to mutate the target
proteins represents an obstacle for effective treatment with molecules
which selectively inhibit the function of specific viral polypeptides.
Thus, there is need for new therapeutic strategies to prevent and treat
viral infections.

[0012]Additionally, there is a need for new therapies for the prevention
and treatment of bacterial infections, especially bacterial infections
caused by multiple drug resistant bacteria. Currently, bacterial
infections are treated with various antibiotics. Although antibiotics
have and can be effective in the treatment of various bacterial
infections, there are a number of limitations to the effectiveness and
safety of antibiotics. For example, some individuals have an allergic
reaction to certain antibiotics and other individuals suffer from serious
side effects. Moreover, continued use of antibiotics for the treatment of
bacterial infections contributes to formation of antibiotic-resistant
strains of bacteria.

[0013]Another aspect of the present invention is directed to the use of
the peptide for prophylaxis and/or treatment of infectious diseases
including opportunistic infections.

[0015]Another aspect of the present invention is directed to the use of
the peptide for prophylaxis and/or treatment of prion diseases.

[0016]Prions are infectious agents which do not have a nucleic acid
genome. It seems that a protein alone is the infectious agent. A prion
has been defined as "small proteinaceous infectious particle which
resists inactivation by procedures that modify nucleic acids". The
discovery that proteins alone can transmit an infectious disease came as
a considerable surprise to the scientific community. Prion diseases are
often called "transmissible spongiform encephalopathies", because of the
post mortem appearance of the brain with large vacuoles in the cortex and
cerebellum.

[0017]Probably most mammalian species develop these diseases. Prion
diseases are a group of neurodegenerative disorders of humans and animals
and the prion diseases can manifest as sporadic, genetic or infectious
disorders. Examples of prion diseases acquired by exogenous infection are
bovine spongiform encephalitis (BSE) of cattle and the new variant of
Creutzfeld-Jakob disease (vCJD) caused by BSE as well as scrapie of
animals. Examples of human prion diseases include kuru, sporadic
Creutzfeldt-Jakob disease (sCJD), familial CJD (fCJD), iatrogenic CJD
(iCJD), Gerstmann-Straussler-Scheinker (GSS) disease, fatal familial
insomnia (FFI), and especially the new variant CJD (nvCJD or vCJD).

[0018]The name "prion" is used to describe the causative agents which
underlie the transmissible spongiform encephalopathies. A prion is
proposed to be a novel infectious particle that differs from viruses and
viroids. It is composed solely of one unique protein that resists most
inactivation procedures such as heat, radiation, and proteases. The
latter characteristic has led to the term protease-resistant isoform of
the prion protein. The protease-resistant isoform has been proposed to
slowly catalyze the conversion of the normal prion protein into the
abnormal form.

[0019]The term "isoform" in the context of prions means two proteins with
exactly the same amino acid sequence that can fold into molecules with
dramatically different tertiary structures. The normal cellular isoform
of the prion protein (PrPc) has a high α-helix content, a low
β-sheet content, and is sensitive to protease digestion. The
abnormal, disease-causing isoform (PrPSc) has a lower α-helix
content, a much higher β-sheet content, and is much more resistant
to protease digestion.

[0021]The peptides of the present invention were tested using the assays
described in Examples 1-7 for their effect as active therapeutic agents
in the prophylaxis and/or treatment of infectious diseases and disorders.

Autoimmune Disease

[0022]Autoimmune disease refers to any of a group of diseases or disorders
in which tissue injury is associated with a humoral and/or cell-mediated
immune response to body constituents or, in a broader sense, an immune
response to self. The pathological immune response may be systemic or
organ specific. That is, for example, the immune response directed to
self may affect joints, skin, myelin sheath that protects neurons,
kidney, liver, pancreas, thyroid, adrenals, and ovaries.

[0023]In fact, the list of autoimmune diseases is composed of more than
eighty disorders. A few autoimmune diseases such as vitiligo, in which
patches of skin lose pigmentation, are merely annoying. Most others are
debilitating, often progressive with time and eventually fatal. Systemic
lupus erythematosus (SLE), for example, is a chronic disease in which
10-15% of patients die within a decade of diagnosis, in all but a few
autoimmune diseases, the sex ratio skews towards women. For example, in
SLE the ratio of female to male patients is nine to one. In one
particular case, Hashimoto's disease in which the immune system attacks
the thyroid gland, the ratio is fifty to one.

[0024]It has long been known that immune complex formation plays a role in
the etiology and progression of autoimmune disease. For example,
inflammation in patients with arthritis has long been considered to
involve phagocytosis by leukocytes of complexes of antigen, antibody and
complement-immune complexes. However, only now it is being recognized
that inflammation caused by immune complexes in the joints (arthritis),
the kidneys (glomerulonephritis), and blood vessels (vasculitis) is a
major cause of morbidity in autoimmune diseases. Increased immune complex
formation correlates with the presence of antibodies directed to self or
so-called autoantibodies, and the presence of the latter can also
contribute to tissue inflammation either as part of an immune complex or
unbound to antigen (free antibody). In some autoimmune diseases, the
presence of free autoantibody contributes significantly to disease
pathology. This has been clearly demonstrated for example in SLE
(anti-DNA antibodies), immune thrombocytopenia (antibody response
directed to platelets), and to a lesser extent rheumatoid arthritis (IgG
reactive rheumatoid factor). The important role of immune complexes and
free autoantibodies is further demonstrated by the fact that successful
treatment of certain autoimmune diseases has been achieved by the removal
of immune complexes and free antibody by means of specific
immunoadsorption procedures. For example, the use of an apheresis
procedure in which immune complexes and antibodies are removed by passage
of a patient's blood through an immunoaffinity column was approved by the
U.S. FDA in 1987 for immune thrombocytopenia (ITP) and in 1999 for
rheumatoid arthritis. However, currently there is no approved method for
the treatment of autoimmune diseases which facilitates the elimination of
immune complexes and autoantibodies by administration of a drug.

[0025]Another aspect of the etiology and progression of autoimmune disease
is the role of proinflammatory cytokines. Under normal circumstances,
proinflammatory cytokines such as tumor necrosis factor α
(TNFα) and interleukin-1 (IL-1) play a protective role in the
response to infection and cellular stress. However, the pathological
consequences which result from chronic and/or excessive production of
TNFα and IL-1 are believed to underlie the progression of many
autoimmune diseases such as rheumatoid arthritis, Crohn's disease,
inflammatory bowel disease, and psoriasis. Other proinflammatory
cytokines include interleukin-6, interleukin-8, interleukin-17, and
granulocyte-macrophage colony stimulating factor.

[0026]Naturally occurring CD4+CD25+ regulatory T cells (Tregs) play a
critical role in the control of periphery tolerance to self-antigens.
Interestingly, they also control immune responses to allergens and
transplant antigens. Recent studies in animal models have shown that
adoptive transfer of CD4+CD25+ Tregs can prevent or even cure allergic
and autoimmune diseases, and appear to induce transplantation tolerance.
Thus, adoptive cell therapy using patient-specific CD4+CD25+ Tregs has
emerged as an individualized medicine for the treatment of inflammatory
disease including allergy, autoimmune disease and transplant rejection.
Furthermore, strategies to activate and expand antigen-specific CD4+CD25+
Tregs in vivo using pharmacological agents may represent a novel avenue
for drug development.

[0027]The interaction of leukocytes with the vessel endothelium to
facilitate the extravasation into the tissue represents a key process of
the body's defense mechanisms. Excessive recruitment of leukocytes into
the inflamed tissue in chronic diseases like autoimmune disorders could
be prevented by interfering with the mechanisms of leukocyte
extravasation. Significant progress in elucidating the molecular basis of
the trafficking of leukocytes from the blood stream to the extravascular
tissue has been achieved that enables new strategies for therapeutic
approaches. The multistep process of leukocyte rolling, firm adhesion and
transmigration through the endothelial wall is facilitated by a dynamic
interplay of adhesion receptors on both leukocytes and on endothelial
cells as well as chemokines. In preclinical studies using various animal
models, promising results have been obtained demonstrating that blocking
of adhesion receptors of the selectin and integrin families improved the
inflammation process in models of ulcerative colitis, autoimmune
encephalomyelitis or contact hypersensitivity. In addition to the
targeting of adhesion receptors by antibodies, small molecules that mimic
epitopes of adhesion receptor ligands have been developed and
successfully applied in animal models. Clinical studies revealed a
limited response using antibodies to selectins or leukocyte
function-associated antigen 1 (LFA-1) integrins compared with animal
models. However, using humanized antibodies to the alpha 4-integrin
subunit significant efficacy has been demonstrated in autoimmune diseases
like psoriasis, multiple sclerosis and inflammatory bowel disease.

[0028]Examples of autoimmune diseases of the eyes are idiopathic
opticus-neuritis, ophthalmia sympathica, anterior uveitis and other
uveitis forms, retina degeneration, and Mooren's ulcer.

[0042]During the last decade, new biotherapies have been developed for the
treatment of systemic autoimmune diseases. The targets of these new
treatments are all the steps of the immune response. These new therapies
are: B lymphocyte (BL) inhibitors such as anti-CD20 monoclonal antibody,
B lymphocyte stimulator (BLyS) antagonists and tolerogens of
pathogenic-antibody secreting LB; inhibitors of the costimulation between
antigen-presenting cells and T lymphocyte (TL) like monoclonal anti-CD40
ligand antibody or CTLA4-Ig (abatecept); TL antagonists which can inhibit
the proliferation of autoreactive T cells; cytokine antagonists;
chemokine and adhesin antagonists which inhibit trafficking of
immunocompetent cells to target organs. These new approaches are based on
a better understanding of the autoimmune response.

[0043]The peptide of the present invention was tested using the assays
described in Examples 14-15 for their effect as active therapeutic agents
in the prophylaxis and/or treatment of autoimmune diseases and disorders.

Fibrotic Disease

[0044]Fibrosis or fibrosis associated disorder affects the liver,
epidermis, endodermis, muscle, tendon, cartilage, heart, pancreas, lung,
uterus, nervous system, testis, ovary, adrenal gland, artery, vein,
colon, small intestine, biliary tract, or stomach. In a further
embodiment, the fibrosis or fibrosis associated disorder is interstitial
lung fibrosis. In another embodiment the fibrosis or fibrosis associated
disorder is the result of an infection with schistosoma. In another
embodiment the fibrosis or fibrosis associated disorder is the result of
wound healing.

[0047]The emergence and disappearance of the myofibroblast appears to
correlate with the initiation of active fibrosis and its resolution,
respectively. In addition, the myofibroblast has many phenotypic
features, which embody much of the pathologic alterations in fibrotic
tissue, e.g. lung tissue. These features would seem to argue for an
important role for the myofibroblast in the pathogenesis of fibrosis,
e.g. lung fibrosis. Furthermore, the persistence of the myofibroblast may
herald progressive disease, and, conversely, its disappearance may be an
indicator of resolution. This in turn suggests that future therapeutic
strategies targeting the myofibroblast would be productive.

[0048]Patients usually exhibit evidence of active fibrosis with increased
numbers of activated fibroblasts, many of which have the phenotypic
characteristics of myofibroblasts. At these sites, increased amounts of
extracellular matrix deposition are evident with effacement of the normal
alveolar architecture. Animal model studies show the myofibroblast to be
the primary source of type I collagen gene expression in active fibrotic
sites. In vitro studies show differentiation of these cells from
fibroblasts under the influence of certain cytokines but indicate their
susceptibility to nitric oxide mediated apoptosis. In addition to
promoting myofibroblast differentiation, transforming growth
factor-β1 (TGF-β1) provides protection against apoptosis. Thus,
this well-known fibrogenic cytokine is important both for the emergence
of the myofibroblast and its survival against apoptotic stimuli. This is
consistent with the critical importance of this cytokine in diverse
models of fibrosis in various tissues. In view of these properties, the
persistence or prolonged survival of the myofibroblast may be the key to
understanding why certain forms of lung injury may result in progressive
disease, terminating in end stage disease.

[0049]Although pulmonary fibrosis has diverse etiologies, there is a
common feature characteristic of this process, namely, the abnormal
deposition of extracellular matrix that effaces the normal lung tissue
architecture. A key cellular source of this matrix is the mesenchymal
cell population that occupies much of the fibrotic lesion during the
active period of fibrosis. This population is heterogeneous with respect
to a number of key phenotypes. One of these phenotypes is the
myofibroblast, which is commonly identified by its expression in
α-smooth muscle actin and by features that are intermediate between
the bona fide smooth muscle cell and the fibroblast. The de novo
appearance of myofibroblasts at sites of wound healing and tissue
repair/fibrosis is associated with the period of active fibrosis and is
considered to be involved in wound contraction. Furthermore, the
localization of myofibroblasts at sites undergoing active extracellular
matrix deposition suggests an important role for these cells in the
genesis of the fibrotic lesion.

Increased TGF-β1 Family Levels in Fibrotic Diseases

[0050]The transforming growth factor-β1 (TGF-β1)
family of proteins has the most potent stimulatory effect on
extracellular matrix deposition of any cytokines so far examined. In
animal models of pulmonary fibrosis enhanced TGF-β1 gene
expression is temporally and spatially related to increased collagen gene
expression and protein deposition. TGF-β1 antibodies reduce
collagen deposition in murine bleomycin-induced lung fibrosis and human
fibrotic lung tissue shows enhanced TGF-β1 gene and protein
expression. Several lines of evidence suggest that TGF-β is a
central regulator of pulmonary fibrosis. Several animal models over
expressing TGF-β showed extensive progressive fibrosis but limited
inflammation, indicating that TGF-β may play a predominant role in
the progression of pulmonary fibrosis. Therapeutic efforts are therefore
focusing on inhibition of TGF-β activity, for instance by
anti-TGF-β1-antibodies, or modulators of TGF-β1 such as
pirfenidone. Pirfenidone inhibits TGF-β1 gene expression in vivo
resulting in inhibition of TGF-β1-mediated collagen synthesis and
appears to slow progression of IPF in patients. Other novel, promising
antifibrotic agents include relaxin (inhibits TGF-β-mediated
overexpression of collagen and increases collagenases), suramin (inhibits
growth factors), prostaglandin E2 (inhibits collagen production) and
lovastatin (blocks formation of granulation tissue by induction of
fibroblast apoptosis).

[0052]An important role of tumor necrosis factor-α (TNF-α) in
interstitial fibrosis has been established using transgenic mice, which
either overexpress or display a deficiency of this cytokine. Mice
transgenically modified to overexpress TNF-α develop lung fibrosis.
In contrast, mice null for TNF-α show marked resistance to
bleomycin induced fibrosis. TNF-α can stimulate fibroblast
replication and collagen synthesis in vitro, and pulmonary TNF-α
gene expression rises after administration of bleomycin in mice. Soluble
TNF-α receptors reduce lung fibrosis in murine models and pulmonary
overexpression of TNF-α in transgenic mice is characterized by lung
fibrosis. In patients with CFA or asbestosis, bronchoalveolar lavage
fluid-derived macrophages release increased amounts of TNF-α
compared with controls.

[0054]The abnormal extracellular matrix (ECM) remodeling observed in the
lungs of patients with interstitial pulmonary fibrosis (IPF) is due, at
least in part, to an imbalance between matrix metalloproteases (MMPs) and
tissue inhibitor of metalloproteinases (TIMPs). Normal lung fibroblasts
do not make MMP-9 in vitro, whereas fibroblasts from IPF lungs strongly
express MMP-9. In addition, fibroblasts from patients with IPF express
increased levels of all TIMPs. In this setting, TIMPs may play a role in
apoptosis in some cell populations. In vitro studies of alveolar
macrophages obtained from untreated patients with idiopathic pulmonary
fibrosis showed marked increase in MMP-9 secretion compared to
macrophages collected from healthy individuals. In animals models of
bleomycin-induced pulmonary fibrosis MMPs have been shown to be elevated
in bronchoalveolar lavage (BAL) fluid. Indeed, a synthetic inhibitor of
MMP, Batimastat, has been shown to significantly reduce bleomycin-induced
lung fibrosis, again pointing to the importance of MMPs in the
development of this fibrotic disease in the lung. A number of studies
have shown that the actions of MMPs can result in the release of growth
factors and cytokines. These profibrotic factors require proteolytic
processing for their activation or release from extracellular matrix or
carrier proteins before they can exert their activity. In fact, the
proteolytic activity processing of several key factors involved in the
pathogenesis of pulmonary fibrosis such as insulin-like growth factor
(IGF), TGF-β1 and TNF-α occur through the actions of
MMPs, thereby activating or releasing them from inhibitory
protein-protein interactions. For example, IGFs in vivo are sequestered
by six high affinity IGF binding proteins (IGFBPs1-6), preventing their
ability to interact with IGF receptors. Studies examining adults and
children IPF and interstitial lung disease show that beside IPF, IGFBP-3
and IFPB-2 levels are increased in IPF BAL fluid. MMPs have recently been
shown to regulate the cleavage of IGF binding proteins, thereby
liberating the complexed ligand to affect IGF actions in target cells.
Observations have also shown that the gelatinases, MMP-9 and MMP-2 may be
involved in proteolytic activation of latent TGF-β complexes.
Furthermore, the MMP inhibitor Batimastat reduces MMP-9 activity in BAL
fluid, which was associated with decreased amount of TGF-β and
TNF-α.

[0055]Pulmonary fibrosis can be an all too common consequence of an acute
inflammatory response of the lung to a host of inciting events. Chronic
lung injury due to fibrotic changes can result from an identifiable
inflammatory event or an insidious, unknown event. The inflammatory
process can include infiltration of various inflammatory cell types, such
as neutrophils and macrophages, the secretion of inflammatory cytokines
and chemokines and the secretion of matrix remodeling proteinases.

[0057]The peptide of the present invention was tested using the assays
described in Examples 14-15 for their effect as active therapeutic agents
in the prophylaxis and/or treatment of fibrotic diseases and disorders.

Inflammatory Disease

[0058]Inflammation is the final common pathway of various insults, such as
infection, trauma, and allergies to the human body. It is characterized
by activation of the immune system with recruitment of inflammatory
cells, production of pro-inflammatory cells and production of
pro-inflammatory cytokines. Most inflammatory diseases and disorders are
characterized by abnormal accumulation of inflammatory cells including
monocytes/macrophages, granulocytes, plasma cells, lymphocytes and
platelets. Along with tissue endothelial cells and fibroblasts, these
inflammatory cells release a complex array of lipids, growth factors,
cytokines and destructive enzymes that cause local tissue damage.

[0059]One form of inflammatory response is neutrophilic inflammation which
is characterized by infiltration of the inflamed tissue by neutrophil
polymorphonuclear leukocytes (PMN), which are a major component of the
host defense. Tissue infection by extracellular bacteria represents the
prototype of this inflammatory response. On the other hand, various
non-infectious diseases are characterized by extravascular recruitment of
neutrophils. This group of inflammatory diseases includes chronic
obstructive pulmonary disease, adult respiratory distress syndrome, some
types of immune-complex alveolitis, cystic fibrosis, bronchitis,
bronchiectasis, emphysema, glomerulonephritis, rheumatoid arthritis,
gouty arthritis, ulcerative colitis, certain dermatoses such as psoriasis
and vasculitis. In these conditions neutrophils are thought to play a
crucial role in the development of tissue injury which, when persistent,
can lead to the irreversible destruction of the normal tissue
architecture with consequent organ dysfunction. Tissue damage is
primarily caused by the activation of neutrophils followed by their
release of proteinases and increased production of oxygen species.

[0060]Chronic obstructive pulmonary disease (COPD) is described by the
progressive development of airflow limitation that is not fully
reversible. Most patients with COPD have three pathological conditions;
bronchitis, emphysema and mucus plugging. This disease is characterized
by a slowly progressive and irreversible decrease in forced expiratory
volume in the first second of expiration (FEVi), with relative
preservation of forced vital capacity (FVC). In both asthma and COPD
there is significant, but distinct, remodeling of airways. Most of the
airflow obstruction is due to two major components, alveolar destruction
(emphysema) and small airways obstruction (chronic obstructive
bronchitis). COPD is mainly characterized by profound mucus cell
hyperplasia. Neutrophil infiltration of the patient's lungs is a primary
characteristic of COPD. Elevated levels of proinflammatory cytokines,
like TNF-α, and especially chemokines like interleukin-8 (IL-8) and
growth-regulated oncogene-α (GRO-α) play a very important
role in pathogenesis of this disease. Platelet thromboxane synthesis is
also enhanced in patients with COPD. Most of the tissue damage is caused
by activation of neutrophils followed by their release of
metalloproteinases, and increased production of oxygen species.

[0061]TNF-α has several biologic activities that are important in
homeostasis as well as in pathophysiological conditions. The main sources
of TNF-α are monocytes-macrophages, T-lymphocytes and mast cells.
The finding that anti-TNF-α antibodies (cA2) are effective in the
treatment of patients suffering from rheumatoid arthritis (RA)
intensified the interest to find new TNF-α inhibitors as possible
potent medicaments for RA. Rheumatoid arthritis is an autoimmune chronic
inflammatory disease characterized by irreversible pathological changes
of the joints. In addition to RA, TNF-α antagonists are also
applicable to several other pathological conditions and diseases such as
spondylitis, osteoarthritis, gout and other arthritic conditions, sepsis,
septic shock, toxic shock syndrome, atopic dermatitis, contact
dermatitis, psoriasis, glomerulonephritis, lupus erythematosus,
scleroderma, asthma, cachexia, chronic obstructive lung disease,
congestive heart failure, insulin resistance, lung (pulmonary) fibrosis,
multiple sclerosis, Crohn's disease, ulcerative colitis, viral infections
and AIDS.

[0064]By "proliferative skin disease" is meant a benign or malignant
disease that is characterized by accelerated cell division in the
epidermis or dermis. Examples of proliferative skin diseases are
psoriasis, atopic dermatitis, nonspecific dermatitis, primary irritant
contact dermatitis, allergic contact dermatitis, basal and squamous cell
carcinomas of the skin, lamellar ichthyosis, epidermolytic
hyperkeratosis, premalignant keratosis, acne, and seborrheic dermatitis.
As will be appreciated by one skilled in the art, a particular disease,
disorder, or condition may be characterized as being both a proliferative
skin disease and an inflammatory dermatosis. An example of such a disease
is psoriasis.

[0085]A human peptide is "active" in an inflammatory disease if the
inhibition is >50% in one of the assays described below. Inhibition
(as percentage) was calculated using the following formula: %
inhibition=(1-concentration of cytokines in sample/concentration of
cytokines in positive control)×100. The positive control refers to
stimulated samples, not treated with substances.

Cyclic AMP Induction

[0086]Adenosine 3', 5'-cyclic monophosphate (cyclic AMP; cAMP) is one of
the most important "second messengers" involved as a modulator of
physiological processes. cAMP is also involved in regulating neuronal,
glandular, cardiovascular, immune and other functions and actions. A
number of hormones are known to activate cAMP through the action of the
enzyme adenylate cyclase, which is located at the cell membranes,
converts ATP to cAMP.

##STR00001##

[0087]Adenylyl cyclase is activated by a range of signaling molecules
through the activation of adenylyl cyclase stimulatory G-protein coupled
receptors.

[0088]cAMP is a second messenger, used for intracellular signal
transduction, such as transferring the effects of peptides like
aviptadil, which cannot get through the cell membrane. Its purposes
include the activation of protein kinases and regulating the effects of
aviptadil or glucagon.

[0089]For being a second messenger, cAMP has the following characteristics
to work effectively: [0090]Amplification--when a few peptide molecules
signal the cell, it needs to respond with a signal that is strong enough
to get the biological response done without repeated high levels of
stimulation. [0091]Control--eventually a response must be down-regulated,
and there are instances when it is necessary for a cell to respond a
little or a lot--so control is needed. [0092]Specificity--when a peptide
is utilized, a given cell should respond in a specific way. Not all cells
will respond to this peptide and therefore it cannot have the same effect
in all the cells.

[0093]cAMP has been shown to be involved in the cardiovascular, nervous
system, and immune mechanisms, cell growth and differentiation, and
general metabolism in a regulatory way.

[0094]Elevated levels of cAMP in human cells are associated with the
suppression of cell activation.

[0096]The peptides of the present invention are useful in the treatment of
a variety of allergic, autoimmune, and inflammatory diseases.

[0097]The term "inflammatory disease," as used herein, means any disease
in which an excessive or unregulated inflammatory response leads to
excessive inflammatory symptoms, host tissue damage, or loss of tissue
function. Additionally, the term "autoimmune disease," as used herein,
means any group of disorders in which tissue injury is associated with
humoral or cell-mediated responses to the body's own constituents. The
term "allergic disease," as used herein, means any symptoms, tissue
damage, or loss of tissue function resulting from allergy. The term
"arthritic disease," as used herein, means any of a large family of
diseases that are characterized by inflammatory lesions of the joints
attributable to a variety of etiologies.

[0098]The present invention also provides a method of modulating cAMP
levels in a mammal, as well as a method of treating diseases
characterized by elevated cytokine levels.

[0099]The term "cytokine," as used herein, means any secreted polypeptide
that affects the functions of other cells, and that modulates
interactions between cells in the immune or inflammatory response.
Cytokines include, but are not limited to monokines, lymphokines, and
chemokines regardless of which cells produce them. For instance, a
monokine is generally referred to as being produced and secreted by a
monocyte, however, many other cells produce monokines, such as natural
killer cells, fibroblasts, basophils, neutrophils, endothelial cells,
brain astrocytes, bone marrow stromal cells, epidermal keratinocytes, and
B-lymphocytes. Lymphokines are generally referred to as being produced by
lymphocyte cells. Examples of cytokines include, but are not limited to,
interleukin-1 (IL-1), interleukin-6 (IL-6), and Tumor Necrosis Factor
alpha.

[0100]The present invention further provides a method of reducing cytokine
levels in a mammal, which comprises administering an effective amount of
the peptides of the invention to the mammal.

[0101]Moreover, the peptides of the present invention are useful in
suppressing inflammatory cell activation. The term "inflammatory cell
activation," as used herein, means the induction by a stimulus
(including, but not limited to, cytokines, antigens or auto-antibodies)
of a proliferative cellular response, the production of soluble mediators
(including but not limited to cytokines, oxygen radicals, enzymes,
prostanoids, or vasoactive amines), or cell surface expression of new or
increased numbers of mediators (including, but not limited to, major
histocompatability antigens or cell adhesion molecules) in inflammatory
cells (including but not limited to monocytes, macrophages, T
lymphocytes, B lymphocytes, granulocytes, polymorphonuclear leukocytes,
mast cells, basophils, eosinophils, dendritic cells, and endothelial
cells).

[0102]The peptides of the present invention also are useful in causing
airway smooth muscle relaxation, bronchodilation, preevention of
bronchoconstriction, and vasodilation in blood vessels.

[0104]Other examples of such diseases or related conditions include
cardiomyopathies, such as congestive heart failure, pyrexia, cachexia,
cachexia secondary to infection or malignancy, cachexia secondary to
acquired immune deficiency syndrome (AIDS), ARC (AIDS-related complex),
cerebral malaria, osteoporosis and bone resorption diseases, and fever
and myalgias due to infection. In addition, the peptides of the present
invention are useful in the treatment of diabetes insipidus and central
nervous system disorders, such as depression and multi-infarct dementia.
Peptides of the present invention also have utility outside of that
typically known as therapeutic. For example, the present compounds can
function as organ transplant preservatives.

[0105]The peptide of the present invention was tested using the assays
described in Examples 1-7, 9-18 for their effect as active therapeutic
agents in the prophylaxis and/or treatment of inflammatory diseases and
disorders.

Neurodegenerative Disease

[0106]The present invention also relates generally to the fields of
neurology and psychiatry and to methods of protecting the cells of a
mammalian central nervous system from damage or injury.

[0107]Injuries or trauma of various kinds to the central nervous system
(CNS) or the peripheral nervous system (PNS) can produce profound and
long-lasting neurological and/or psychiatric symptoms and disorders. One
form that this can take is the progressive death of neurons or other
cells of the central nervous system (CNS), i.e., neurodegeneration or
neuronal degeneration.

[0108]Neuronal degeneration as a result of, for example; Alzheimer's
disease, multiple sclerosis, cerebral-vascular accidents (CVAs)/stroke,
traumatic brain injury, spinal cord injuries, degeneration of the optic
nerve, e.g., ischemic optic neuropathy or retinal degeneration and other
central nervous system disorders is an enormous medical and public health
problem by virtue of both its high incidence and the frequency of
long-term sequelae. Animal studies and clinical trials have shown that
amino acid transmitters (especially glutamate), oxidative stress and
inflammatory reactions contribute strongly to cell death in these
conditions. Upon injury or upon ischemic insult, damaged neurons release
massive amounts of the neurotransmitter glutamate, which is excitotoxic
to the surrounding neurons. Glutamate is a negatively charged amino acid
that is an excitatory synaptic transmitter in the mammalian nervous
system. Although the concentration of glutamate can reach the millimolar
range in nerve terminals its extracellular concentration is maintained at
a low level to prevent neurotoxicity. It has been noted that glutamate
can be toxic to neurons if presented at a high concentration. The term
"excitotoxicity" has been used to describe the cytotoxic effect that
glutamate (and other such excitatory amino acids) can have on neurons
when applied at high dosages.

[0109]Patients with injury or damage of any kind to the central (CNS) or
peripheral (PNS) nervous system including the retina may benefit from
neuroprotective methods. This nervous system injury may take the form of
an abrupt insult or an acute injury to the nervous system as in, for
example, acute neurodegenerative disorders including, but not limited to;
acute injury, hypoxia-ischemia or the combination thereof resulting in
neuronal cell death or compromise. Acute injury includes, but is not
limited to, traumatic brain injury (TBI) including, closed, blunt or
penetrating brain trauma, focal brain trauma, diffuse brain damage,
spinal cord injury, intracranial or intravertebral lesions (including,
but not limited to, contusion, penetration, shear, compression or
laceration lesions of the spinal cord or whiplash shaken infant
syndrome).

[0112]In addition, trauma and progressive injury to the nervous system can
take place in various psychiatric disorders, including but not limited
to, progressive, deteriorating forms of bipolar disorder or
schizoaffective disorder or schizophrenia, impulse control disorders,
obsessive compulsive disorder (OCD), behavioral changes in temporal lobe
epilepsy and personality disorders.

[0113]In one preferred embodiment the compounds of the invention would be
used to provide neuroprotection in disorders involving trauma and
progressive injury to the nervous system in various psychiatric
disorders. These disorders would be selected from the group consisting
of; schizoaffective disorder, schizophrenia, impulse control disorders,
obsessive compulsive disorder (OCD) and personality disorders.

[0114]In addition, trauma and injury make take the form of disorders
associated with overt and extensive memory loss including, but not
limited to, neurodegenerative disorders associated with age-related
dementia, vascular dementia, diffuse white matter disease (Binswanger's
disease), dementia of endocrine or metabolic origin, dementia of head
trauma and diffuse brain damage, dementia pugilistica or frontal lobe
dementia, including but not limited to Pick's Disease.

[0120]The term "neuroprotection" as used herein shall mean; inhibiting,
preventing, ameliorating or reducing the severity of the dysfunction,
degeneration or death of nerve cells, axons or their supporting cells in
the central or peripheral nervous system of a mammal, including a human.
This includes the treatment or prophylaxis of a neurodegenerative
disease; protection against excitotoxicity or ameliorating the cytotoxic
effect of a compound (for example, a excitatory amino acid such as
glutamate; a toxin; or a prophylactic or therapeutic compound that exerts
an immediate or delayed cytotoxic side effect including but not limited
to the immediate or delayed induction of apoptosis) in a patient in need
thereof.

[0121]The term "a patient in need of treatment with a neuroprotective
drug" as used herein will refer to any patient who currently has or may
develop any of the above syndromes or disorders, or any disorder in which
the patient's present clinical condition or prognosis could benefit from
providing neuroprotection to prevent the development, extension,
worsening or increased resistance to treatment of any neurological or
psychiatric disorder.

[0122]The term "treating" or "treatment" as used herein, refers to any
indicia of success in the prevention or amelioration of an injury,
pathology or condition, including any objective or subjective parameter
such as abatement; remission; diminishing of symptoms or making the
injury, pathology, or condition more tolerable to the patient; slowing in
the rate of degeneration or decline; making the final point of
degeneration less debilitating; or improving a subject's physical or
mental well-being. The treatment or amelioration of symptoms can be based
on objective or subjective parameters; including the results of a
physical examination, neurological examination, and/or psychiatric
evaluations.

[0123]In some embodiments this invention provides methods of
neuroprotection. In certain embodiments, these methods comprise
administering a therapeutically effective amount of the peptide of the
invention to a patient who has not yet developed overt, clinical signs or
symptoms of injury or damage to the cells of the nervous system but who
may be in a high risk group for the development of neuronal damage
because of injury or trauma to the nervous system or because of some
known predisposition either biochemical or genetic or the finding of a
verified biomarker of one or more of these disorders.

[0124]Thus, in some embodiments, the methods and compositions of the
present invention are directed toward neuroprotection in a subject who is
at risk of developing neuronal damage but who has not yet developed
clinical evidence. This patient may simply be at "greater risk" as
determined by the recognition of any factor in a subject's, or their
families, medical history, physical exam or testing that is indicative of
a greater than average risk for developing neuronal damage. Therefore,
this determination that a patient may be at a "greater risk" by any
available means can be used to determine whether the patient should be
treated with the methods of the present invention.

[0125]Accordingly, in an exemplary embodiment, subjects who may benefit
from treatment by the methods and peptide of this invention can be
identified using accepted screening methods to determine risk factors for
neuronal damage. These screening methods include, for example,
conventional work-ups to determine risk factors including but not limited
to: for example, head trauma, either closed or penetrating, CNS
infections, bacterial or viral, cerebrovascular disease including but not
limited to stroke, brain tumors, brain edema, cysticercosis, porphyria,
metabolic encephalopathy, drug withdrawal including but not limited to
sedative-hypnotic or alcohol withdrawal, abnormal perinatal history
including anoxia at birth or birth injury of any kind, cerebral palsy,
learning disabilities, hyperactivity, history of febrile convulsions as a
child, history of status epilepticus, family history of epilepsy or any
seizure related disorder, inflammatory disease of the brain including
lupis, drug intoxication either direct or by placental transfer,
including but not limited to cocaine poisoning, parental consanguinity,
and treatment with medications that are toxic to the nervous system
including psychotropic medications.

[0126]The determination of which patients may benefit from treatment with
a neuroprotective drug in patients who have no clinical signs or symptoms
may be based on a variety of "surrogate markers" or "biomarkers".

[0127]As used herein, the terms "surrogate marker" and "biomarker" are
used interchangeably and refer to any anatomical, biochemical,
structural, electrical, genetic or chemical indicator or marker that can
be reliably correlated with the present existence or future development
of neuronal damage. In some instances, brain-imaging techniques, such as
computer tomography (CT), magnetic resonance imaging (MRI) or positron
emission tomography (PET), can be used to determine whether a subject is
at risk for neuronal damage. Suitable biomarkers for the methods of this
invention include, but are not limited to: the determination by MRI, CT
or other imaging techniques, of sclerosis, atrophy or volume loss in the
hippocampus or overt mesial temporal sclerosis (MTS) or similar relevant
anatomical pathology; the detection in the patient's blood, serum or
tissues of a molecular species such as a protein or other biochemical
biomarker, e.g., elevated levels of ciliary neurotrophic factor (CNTF) or
elevated serum levels of a neuronal degradation product; or other
evidence from surrogate markers or biomarkers that the patient is in need
of treatment with a neuroprotective drug.

[0128]It is expected that many more such biomarkers utilizing a wide
variety of detection techniques will be developed in the future. It is
intended that any such marker or indicator of the existence or possible
future development of neuronal damage, as the latter term is used herein,
may be used in the methods of this invention for determining the need for
treatment with the compounds and methods of this invention.

[0129]A determination that a subject has, or may be at risk for
developing, neuronal damage would also include, for example, a medical
evaluation that includes a thorough history, a physical examination, and
a series of relevant bloods tests. It can also include an
electroencephalogram (EEG), CT, MRI or PET scan. A determination of an
increased risk of developing neuronal damage or injury may also be made
by means of genetic testing, including gene expression profiling or
proteomic techniques. For psychiatric disorders that may be stabilized or
improved by a neuroprotective drug, e.g., bipolar disorder,
schizoaffective disorder, schizophrenia, impulse control disorders, etc.
the above tests may also include a present state exam and a detailed
history of the course of the patients symptoms such as mood disorder
symptoms and psychotic symptoms over time and in relation to other
treatments the patient may have received over time, e.g., a life chart.
These and other specialized and routine methods allow the clinician to
select patients in need of therapy using the methods and formulations of
this invention. In some embodiments of the present invention peptide
suitable for use in the practice of this invention will be administered
either singly or concomitantly with at least one or more other compounds
or therapeutic agents, e.g., with other neuroprotective drugs or
antiepileptic drugs, anticonvulsant drugs. In these embodiments, the
present invention provides methods to treat or prevent neuronal injury in
a patient. The method includes the step of; administering to a patient in
need of treatment, an effective amount of one of the peptide disclosed
herein in combination with an effective amount of one or more other
compounds or therapeutic agents that have the ability to provide
neuroprotection or to treat or prevent seizures or epileptogenesis or the
ability to augment the neuroprotective effects of the compounds of the
invention.

[0130]As used herein the term "combination administration" of a compound,
therapeutic agent or known drug with the peptide of the present invention
means administration of the drug and the one or more compounds at such
time that both the known drug and the peptide will have a therapeutic
effect. In some cases this therapeutic effect will be synergistic. Such
concomitant administration can involve concurrent (i.e. at the same
time), prior, or subsequent administration of the drug with respect to
the administration of the peptide of the present invention. A person of
ordinary skill in the art would have no difficulty determining the
appropriate timing, sequence and dosages of administration for particular
drugs and peptides of the present invention.

[0131]The said one or more other compounds or therapeutic agents may be
selected from compounds that have one or more of the following
properties: antioxidant activity; NMDA receptor antagonist activity,
augmentation of endogenous GABA inhibition; NO synthase inhibitor
activity; iron binding ability, e.g., an iron chelator; calcium binding
ability, e.g., a Ca (II) chelator; zinc binding ability, e.g., a Zn (II)
chelator; the ability to effectively block sodium or calcium ion
channels, or to open potassium or chloride ion channels in the CNS of a
patient.

[0132]The peptide of the present invention was tested using the assays
described in Examples 1-7, 9-18 for their effect as active therapeutic
agents in the prophylaxis and/or treatment of neurodegenerative diseases
and disorders.

Heart and Vascular Disease

[0133]Heart disease is a general term used to describe many different
heart conditions. For example, coronary artery disease, which is the most
common heart disease, is characterized by constriction or narrowing of
the arteries supplying the heart with oxygen-rich blood, and can lead to
myocardial infarction, which is the death of a portion of the heart
muscle. Heart failure is a condition resulting from the inability of the
heart to pump an adequate amount of blood through the body. Heart failure
is not a sudden, abrupt stop of heart activity but, rather, typically
develops slowly over many years, as the heart gradually loses its ability
to pump blood efficiently. Risk factors for heart failure include
coronary artery disease, hypertension, valvular heart disease,
cardiomyopathy, disease of the heart muscle, obesity, diabetes, and/or a
family history of heart failure.

[0135]Vascular diseases are often the result of decreased perfusion in the
vascular system or physical or biochemical injury to the blood vessel.

[0136]Peripheral vascular disease (PVD) is defined as a disease of blood
vessels often encountered as narrowing of the vessels of the limbs. There
are two main types of these disorders, functional disease which doesn't
involve defects in the blood vessels but rather arises from stimuli such
as cold, stress, or smoking, and organic disease which arises from
structural defects in the vasculature such as atherosclerotic lesions,
local inflammation, or traumatic injury. This can lead to occlusion of
the vessel, aberrant blood flow, and ultimately to tissue ischemia.

[0137]One of the more clinically significant forms of PVD is peripheral
artery disease (PAD). PAD is often treated by angioplasty and
implantation of a stent or by artery bypass surgery. Clinical
presentation depends on the location of the occluded vessel. For example,
narrowing of the artery that supplies blood to the intestine can result
in severe postprandial pain in the lower abdomen resulting from the
inability of the occluded vessel to meet the increased oxygen demand
arising from digestive and absorptive processes. In severe forms the
ischemia can lead to intestinal necrosis. Similarly, PAD in the leg can
lead to intermittent pain, usually in the calf, that comes and goes with
activity. This disorder is known as intermittent claudication (IC) and
can progress to persistent pain while resting, ischemic ulceration, and
even amputation.

[0138]Peripheral vascular disease is also manifested in atherosclerotic
stenosis of the renal artery, which can lead to renal ischemia and kidney
dysfunction.

[0139]One disease in which vascular diseases and their complications are
very common is diabetes mellitus. Diabetes mellitus causes a variety of
physiological and anatomical irregularities, the most prominent of which
is the inability of the body to utilize glucose normally, which results
in hyperglycemia. Chronic diabetes can lead to complications of the
vascular system which include atherosclerosis, abnormalities involving
large and medium size blood vessels (macroangiopathy) and abnormalities
involving small blood vessels (microangiopathy) such as arterioles and
capillaries.

[0140]Patients with diabetes mellitus are at increased risk of developing
one or more foot ulcers as a result of established long-term
complications of the disease, which include impaired nerve function
(neuropathy) and/or ischemia. Local tissue ischemia is a key contributing
factor to diabetic foot ulceration.

[0141]In addition to large vessel disease, patients with diabetes suffer
further threat to their skin perfusion in at least two additional ways.
First, by involvement of the non-conduit arteries, which are
detrimentally affected by the process of atherosclerosis, and secondly,
and perhaps more importantly, by impairment of the microcirculatory
control mechanisms (small vessel disease). Normally, when a body part
suffers some form of trauma, the body part will, as part of the body's
healing mechanism, experience an increased blood flow. When small vessel
disease and ischemia are both present, as in the case of many diabetics,
this natural increased blood flow response is significantly reduced. This
fact, together with the tendency of diabetics to form blood clots
(thrombosis) in the microcirculatory system during low levels of blood
flow, is believed to be an important factor in ulcer pathogenesis.

[0142]Neuropathy is a general term which describes a disease process which
leads to the dysfunction of the nervous system, and is one of the major
complications of diabetes mellitus, with no well-established therapies
for either its symptomatic treatment or for prevention of progressive
decline in nerve function.

[0143]The thickening and leakage of capillaries caused by diabetes
primarily affect the eyes (retinopathy) and kidneys (nephropathy). The
thickening and leakage of capillaries caused by diabetes are also
associated with skin disorders and disorders of the nervous system
(neuropathy).

[0145]Other diseases, although not known to be related to diabetes are
similar in their physiological effects on the peripheral vascular system.
Such diseases include Raynaud syndrome, CREST syndrome, autoimmune
diseases such as erythematosis, rheumatoid disease, and the like.

[0147]The peptide of the present invention was tested using the assays
described in Examples 1-7, 9-18 for their effect as active therapeutic
agents in the prophylaxis and/or treatment of heart and vascular diseases
and disorders.

Rare or Orphan Diseases

[0148]Another aspect of the present invention is directed to the use of
the peptide as a therapeutic agent for the prophylaxis and/or treatment
of a heart and vascular disease, an autoimmune disease, a fibrotic
disease, an inflammatory disease, a neurodegenerative disease, or an
infectious disease, in patients suffering from one or more of the
following Rare or Orphan Diseases:

[0149]Still another aspect of the present invention relates to the use of
the peptide according to claim 1 as an active ingredient, together with
at least one pharmaceutically acceptable carrier, excipient and/or
diluents for the manufacture of a pharmaceutical composition for the
treatment and/or prophylaxis of cancer, an autoimmune disease, a fibrotic
disease, an inflammatory disease, a neurodegenerative disease, an
infectious disease, a lung disease, a heart and vascular disease or a
metabolic disease or any other disease disclosed herein.

[0150]Such pharmaceutical compositions comprise the peptide as an active
ingredient, together with at least one pharmaceutically acceptable
carrier, excipient, binders, disintegrates, glidents, diluents,
lubricants, coloring agents, sweetening agents, flavoring agents,
preservatives or the like. The pharmaceutical compositions of the present
invention can be prepared in a conventional solid or liquid carrier or
diluents and a conventional pharmaceutically-made adjuvant at suitable
dosage level in a known way.

[0151]Preferably the peptide is suitable for intravenous administration or
suitable for oral administration or suitable for administration by
inhalation.

[0153]The present invention also includes the mammalian milk, artificial
mammalian milk as well as mammalian milk substitutes as a formulation for
oral administration of the peptide to newborns, toddlers, and infants,
either as pharmaceutical preparations, and/or as dietary food
supplements.

[0154]The peptide of the invention can also be administered in form of its
pharmaceutically active salts. Suitable pharmaceutically active salts
comprise acid addition salts and alkali or earth alkali salts. For
instance, sodium, potassium, lithium, magnesium or calcium salts can be
obtained.

[0156]The pharmaceutical compositions according to the present invention
will typically be administered together with suitable carrier materials
selected with respect to the intended form of administration, i.e. for
oral administration in the form of tablets, capsules (either solid
filled, semi-solid filled or liquid filled), powders for constitution,
aerosol preparations consistent with conventional pharmaceutical
practices. Other suitable formulations are gels, elixirs, dispersible
granules, syrups, suspensions, creams, lotions, solutions, emulsions,
suspensions, dispersions, and the like. Suitable dosage forms for
sustained release include tablets having layers of varying disintegration
rates or controlled release polymeric matrices impregnated with the
active components and shaped in tablet form or capsules containing such
impregnated or encapsulated porous polymeric matrices. The pharmaceutical
compositions may be comprised of 5 to 95% by weight of the peptide.

[0158]Suitable binders include starch, gelatin, natural sugars, corn
sweeteners, natural and synthetic gums such as acacia, sodium alginate,
carboxymethyl-cellulose, polyethylene glycol and waxes. Among the
lubricants that may be mentioned for use in these dosage forms, boric
acid, sodium benzoate, sodium acetate, sodium chloride, and the like.
Disintegrants include starch, methylcellulose, guar gum and the like.
Sweetening and flavoring agents and preservatives may also be included
where appropriate. Some of the terms noted above, namely disintegrants,
diluents, lubricants, binders and the like, are discussed in more detail
below.

[0159]Additionally, the compositions of the present invention may be
formulated in sustained release form to provide the rate controlled
release of any one or more of the components or active ingredients to
optimize the therapeutic effects. Suitable dosage forms for sustained
release include layered tablets containing layers of varying
disintegration rates or controlled release polymeric matrices impregnated
with the active components and shaped in tablet form or capsules
containing such impregnated or encapsulated porous polymeric matrices.

[0160]Aerosol preparations suitable for inhalation may include solutions
and solids in powder form, which may be in combination with a
pharmaceutically acceptable carrier such as inert compressed gas, e.g.
nitrogen.

[0161]For preparing suppositories, a low melting wax such as a mixture of
fatty acid glycerides such as cocoa butter is first melted, and the
active ingredient is dispersed homogeneously therein by stirring or
similar mixing. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool and thereby solidify.

[0162]Also included are solid form preparations which are intended to be
converted, shortly before use, to liquid form preparations for either
oral or parenteral administration. Such liquid forms include solutions,
suspensions and emulsions.

[0163]The peptide of the present invention may also be deliverable
transdermally. The transdermal compositions may take the form of creams,
lotions, aerosols and/or emulsions and can be included in a transdermal
patch of the matrix or reservoir type as are conventional in the art for
this purpose.

[0164]The transdermal formulation of the peptide of the invention is
understood to increase the bioavailability of said peptide into the
circulating blood. One problem in the administration of peptides is the
loss of bioactivity due to the formation of insolubles in aqueous
environments or due to degradation. Therefore stabilization of peptides
for maintaining their fluidity and maintaining their biological activity
upon administration to the patients in need thereof needs to be achieved.

[0165]Prior efforts to provide active agents for medication include
incorporating the medication in a polymeric matrix whereby the active
ingredient is released into the systemic circulation. Known
sustained-release delivery means of active agents are disclosed, for
example, in U.S. Pat. No. 4,235,988, U.S. Pat. No. 4,188,373, U.S. Pat.
No. 4,100,271, U.S. Pat. No. 4,474,71, U.S. Pat. No. 4,474,752, U.S. Pat.
No. 4,474,753, or U.S. Pat. No. 4,478,822 relating to polymeric
pharmaceutical vehicles for delivery of pharmaceutically active chemical
materials to mucous membranes. The pharmaceutical carriers are aqueous
solutions of certain polyoxyethylene-polyoxypropylene condensates. These
polymeric pharmaceutical vehicles are described as providing for
increased drug absorbtion by the mucous membrane and prolonged drug
action by a factor of two or more. The substituents are block copolymers
of polyoxypropylene and polyoxyethylene used for stabilization of drugs
such as insulin.

[0166]Aqueous solutions of polyoxyethylene-polyoxypropylene block
copolymers (poloxamers) are useful as stabilizers for the peptide. Aside
from serving as a stabilizer for the peptide, poloxamers provide
excellent vehicles for the delivery of the peptide, and they are
physiologically acceptable. Poloxamers, also known by the trade name
Pluronics (e.g. Pluronic F127, Pluronic P85, Pluronic F68) have
surfactant properties that make them useful in industrial applications.
Among other things, they can be used to increase the water solubility of
hydrophobic, oily substances or otherwise increase the miscibility of two
substances with different hydrophobicities. For this reason, these
polymers are commonly used in industrial applications, cosmetics, and
pharmaceuticals. They have also been used as model systems for drug
delivery applications. In situ gelation of pharmaceutical compositions
based on poloxamer that are biologically triggered are known in the art
(e.g. U.S. Pat. No. 5,256,396), describing compositions containing
poloxamer 407 and water at specified concentrations.

[0167]The term capsule refers to a special container or enclosure made of
methyl cellulose, polyvinyl alcohols, or denatured gelatins or starch for
holding or containing compositions comprising the active ingredients.
Hard shell capsules are typically made of blends of relatively high gel
strength bone and pork skin gelatins. The capsule itself may contain
small amounts of dyes, opaquing agents, plasticizers and preservatives.

[0168]Tablet means compressed or molded solid dosage form containing the
active ingredients with suitable diluents. The tablet can be prepared by
compression of mixtures or granulations obtained by wet granulation, dry
granulation or by compaction well known to a person skilled in the art.

[0169]Oral gels refers to the active ingredients dispersed or solubilized
in a hydrophilic semi-solid matrix.

[0170]Powders for constitution refer to powder blends containing the
active ingredients and suitable diluents which can be suspended in water
or juices. One example for such an oral administration form for newborns,
toddlers and/or infants is a human breast milk substitute which is
produced from milk powder and milk whey powder, optionally and partially
substituted with lactose.

[0171]Human breast milk is a complex fluid, rich in nutrients and in
non-nutritional bioactive components. It contains all of the nutrients
needed by the newborn baby. These include the metabolic components (fat,
protein, and carbohydrates), water, and the raw materials for tissue
growth and development, such as fatty acids, amino acids, minerals,
vitamins, and trace elements.

[0172]More than 98% of the fat in is in the form of triglycerides. Oleic
acid and palmitic acid are the most abundant fatty acids in breastmilk
triglycerides, with comparatively high proportions of the essential fatty
acids, and linolenic acid, followed by long-chain polyunsaturated fatty
acids, such as arachidonic acid and docosahexaenoic acid. These
long-chain fatty acids are constituents of brain and neural tissue and
are needed in early life for mental and visual development. The lipid
component of breast milk is the transport vehicle for fat-soluble
micronutrients such as prostaglandins and vitamins A, D, E, and K.

[0173]Proteins account for approximately 75% of the nitrogen-containing
compounds in breast milk. Non-protein nitrogen substances include urea,
nucleotides, peptides, free amino acids, and DNA. The proteins of breast
milk can be divided into two categories: micellar caseins and aqueous
whey proteins, present in the ratio of about 40:60. Casein forms micelles
of relatively small volume and produces a soft, flocculent curd in the
infant's stomach. The major whey proteins are lactalbumin, lactoferrin,
secretory IgA, and serum albumin, with a large number of other proteins
and peptides present in smaller amounts.

[0174]The principal carbohydrate is lactose, a disaccharide produced in
the mammary epithelial cell from glucose by a reaction involving
lactalbumin.

[0175]In addition to the nutritional components, breast milk contains a
wealth of bioactive components that have beneficial non-nutritional
functions. These include a wide range of specific and non-specific
antimicrobial factors; cytokines and anti-inflammatory substances; and
hormones, growth modulators, and digestive enzymes (Table 1), many of
which have multiple activities. These components may be of particular
importance for young infants because of the immaturity of the host
defense and digestive systems early in life.

[0176]Besides breast milk, infant formula is the only other infant milk
which the medical community considers nutritionally acceptable for
infants under the age of one year. Cow's milk is not recommended because
of its high protein and electrolyte (salt) content which may harm
infant's immature kidneys. The nutrient content of infant formula should
comprise: Protein, Fat, Linoleic acid, Vitamins: A, C, D, E, K, thiamin
(B1), riboflavin (B2), B6, B12, Niacin, Folic acid, Pantothenic acid,
Calcium, Metals: magnesium, iron, zinc, manganese, copper; Phosphorus,
Iodine, Sodium chloride, Potassium chloride. In addition, formulas not
made with cow's milk must include biotin, choline, and inositol.
Hypoallergenic formulas reduce the likelihood of certain medical
complications in babies with specific health problems. Baby formula can
be synthesized from raw amino acids. This kind of formula is sometimes
referred to as elemental infant formula or as medical food because of its
specialized nature. Powder blends containing the active ingredients and
suitable diluents which can be suspended in water or juices can be
produced by spray drying.

[0177]Spray drying has been found the most suitable process for removing
the last part of the water, since spray drying can convert milk
concentrate into a powder while still keeping the valuable properties of
the milk. The principle of all spray dryers is to transform the
concentrate into many small droplets which are then exposed to a fast
current of hot air. Because of the very large surface area of the
droplets, the water evaporates almost instantaneously and the droplets
are transformed into powder particles.

[0178]Powdered milk is a powder made from dried milk solids. Powdered milk
has a far longer shelf life than liquid milk and does not need to be
refrigerated due to its low moisture content.

[0179]Instant milk powder is produced by partially rehydrating the dried
milk powder particles causing them to become sticky and agglomerate. The
water is then removed by drying resulting in an increased amount of air
incorporated between the powder particles.

[0180]Milk powder manufacture is a process carried out on a large scale.
It involves the gentle removal of water, while retaining all the
desirable natural properties of the milk like colour, flavour,
solubility, nutritional value.

[0182]The artificial mother milk formulations or mother milk substitutes
of the present invention are preferably prepared by adding to a mother
milk formulation including commercially available mother milk
formulations especially in power form the peptide of the present
invention. The peptide is preferably added in an amount of 3-100 μg
peptide or per 100 ml (commercially available) mother milk formulation,
more preferably in an amount of 5-70 μg/100 ml and most preferably in
an amount of 10-40 μg/100 ml mother milk formulation.

[0183]Suitable diluents are substances that usually make up the major
portion of the composition or dosage form. Suitable diluents include
sugars such as lactose, sucrose, mannitol and sorbitol, starches derived
from wheat, corn rice and potato, and celluloses such as microcrystalline
cellulose. The amount of diluents in the composition can range from about
5 to about 95% by weight of the total composition, preferably from about
25 to about 75%, more preferably from about 30 to about 60% by weight,
and most preferably from about 40 to 50% by weight.

[0184]The term disintegrants refers to materials added to the composition
to help it break apart (disintegrate) and release the medicaments.
Suitable disintegrants include starches, "cold water soluble" modified
starches such as sodium carboxymethyl starch, natural and synthetic gums
such as locust bean, karaya, guar, tragacanth and agar, cellulose
derivatives such as methylcellulose and sodium carboxymethylcellulose,
microcrystalline celluloses and cross-linked microcrystalline celluloses
such as sodium croscarmellose, alginates such as alginic acid and sodium
alginate, clays such as bentonites, and effervescent mixtures. The amount
of disintegrant in the composition can range from about 1 to about 40% by
weight of the composition, preferably 2 to about 30% by weight of the
composition, more preferably from about 3 to 20% by weight of the
composition, and most preferably from about 5 to about 10% by weight.

[0185]Binders characterize substances that bind or "glue" powders together
and make them cohesive by forming granules, thus serving as the
"adhesive" in the formulation. Binders add cohesive strength already
available in the diluents or bulking agent. Suitable binders include
sugars such as sucrose, starches derived from wheat, corn rice and
potato; natural gums such as acacia, gelatin and tragacanth; derivatives
of seaweed such as alginic acid, sodium alginate and ammonium calcium
alginate; cellulosic materials such as methylcellulose and sodium
carboxymethylcellulose and hydroxypropyl-methylcellulose;
polyvinylpyrrolidone; and inorganics such as magnesium aluminum silicate.
The amount of binder in the composition can range from about 1 to 30% by
weight of the composition, preferably from about 2 to about 20% by weight
of the composition, more preferably from about 3 to about 10% by weight,
even more preferably from about 3 to about 6% by weight.

[0186]Lubricant refers to a substance added to the dosage form to enable
the tablet, granules, etc. after it has been compressed, to release from
the mold or die by reducing friction or wear. Suitable lubricants include
metallic stearates such as magnesium stearate, calcium stearate or
potassium stearate; stearic acid; high melting point waxes; and water
soluble lubricants such as sodium chloride, sodium benzoate, sodium
acetate, sodium oleate, polyethylene glycols and d'l-leucine. Lubricants
are usually added at the very last step before compression, since they
must be present on the surfaces of the granules and in between them and
the parts of the tablet press. The amount of lubricant in the composition
can range from about 0.05 to about 15% by weight of the composition,
preferably 0.2 to about 5% by weight of the composition, more preferably
from about 0.3 to about 3%, and most preferably from about 0.3 to about
1.5% by weight of the composition.

[0187]Glidents are materials that prevent caking and improve the flow
characteristics of granulations, so that flow is smooth and uniform.
Suitable glidents include silicon dioxide and talc. The amount of glident
in the composition can range from about 0.01 to 10% by weight of the
composition, preferably 0.1% to about 7% by weight of the total
composition, more preferably from about 0.2 to 5% by weight, and most
preferably from about 0.5 to about 2% by weight.

[0188]Coloring agents are excipients that provide coloration to the
composition or the dosage form. Such excipients can include food grade
dyes and food grade dyes adsorbed onto a suitable adsorbent such as clay
or aluminum oxide. The amount of the coloring agent can vary from about
0.01 to 10% by weight of the composition, preferably from about 0.05 to
6% by weight, more preferably from about 0.1 to about 4% by weight of the
composition, and most preferably from about 0.1 to about 1%.

[0189]The peptide of the invention can be used to form multiparticulates,
discrete particles, well known dosage forms, whose totality represents
the intended therapeutically useful dose of a drug. When taken orally,
multiparticulates generally disperse freely in the gastrointestinal
tract, and maximize absorption. A specific example is described in U.S.
Pat. No. 6,068,859, disclosing multiparticulates that provide controlled
release of azithromycin. Another advantage of the multiparticulates is
the improved stability of the drug. The poloxamer component of the
multiparticulate is very inert, thus minimizing degradation of the drug.

[0190]However, formulation problems result from the melt-congeal process
often used to form multiparticulates. The multiparticulates are
preferably formed into round beads or spheres. Some carriers, when melted
and then solidified, do not form round beads but may solidify into rods,
strings, or other non-spherical shapes. The result is very irregularly
shaped multiparticulates that are difficult to process into dosage forms.
This problem is solved by e.g. WO 2007104173 where the particles consist
of a poloxamer, a resin, and/or a tocopherol, creating together with the
medicament (e.g. insulin) micelles. Micelle formation is essential for
the absorption of many nutrients within the human body. Bile salts formed
in the liver and secreted by the gall bladder allow micelles of fatty
acids to form. This allows the absorption of complicated lipids and lipid
soluble vitamins within the micelle by the small intestine. Micelles are
approximately spherical in shape. Preferably, peptide of the invention
are formulated with a poloxamer and a resin to form micelles suitable for
oral administration to patients in need of the medicament.

[0191]Liquid form preparations include solutions, suspensions and
emulsions. As an example may be mentioned water or water-propylene glycol
solutions for parenteral injections or addition of sweeteners and
opacifiers for oral solutions, suspensions and emulsions. Liquid form
preparations may also include solutions for intranasal administration.

[0193]Preferred is the group of carboxylic acid buffers such as acetate
and carboxylic diacid buffers such as fumarate, tartrate and phthalate
and carboxylic triacid buffers such as citrate. Another group of
preferred buffers is represented by inorganic buffers such as sulfate,
borate, carbonate, oxalate, calcium hydroxyde and phosphate buffers.
Another group of preferred buffers are nitrogen containing buffers such
as imidazole, diethylenediamine, and piperazine.

[0195]Another group of preferred buffers are glycine buffers such as
glycine, glycyl-glycine, glycyl-glycyl-glycine,
N,N-bis(2-hydroxyethyl)glycine and
N-[2-hydroxy-1,1-bis(hydroxy-methyl)ethyl]glycine (Tricine).

[0197]Preferred are the buffers having an effective pH range of from 2.7
to 8.5, and more preferred of from 3.8 to 7.7. The effective pH range for
each buffer can be defined as pKa-1 to pKa+1, where Ka is the ionization
constant for the weak acid in the buffer and pKa=-log K.

[0198]Most preferred are buffers suitable for pharmaceutical use e.g.
buffers suitable for administration to a patient such as acetate,
carbonate, citrate, fumarate, glutamate, lactate, phosphate, phthalate,
and succinate buffers. Particularly preferred examples of commonly used
pharmaceutical buffers are acetate buffer, citrate buffer, glutamate
buffer and phosphate buffer. Also most preferred is the group of
carboxylic acid buffers. The term "carboxylic acid buffers" as used
herein shall refer to carboxylic mono acid buffers and carboxylic diacid
buffers as well as carboxylic triacid buffers. Of course also
combinations of buffers, especially of the buffers mentioned herein are
useful for the present invention.

[0199]Some suitable pharmaceutical buffers are a citrate buffer
(preferably at a final formulation concentration of from about 20 to 200
mM, more preferably at a final concentration of from about 30 to 120 mM)
or an acetate buffer (preferably at a final formulation concentration of
about 20 to 200 mM) or a phosphate buffer (preferably at a final
formulation concentration of about 20 to 200 mM).

[0200]Techniques for the formulation and administration of the peptide of
the present invention may be found in "Remington's Pharmaceutical
Sciences" Mack Publishing Co., Easton Pa. A suitable composition
comprising the peptide mentioned herein may be a solution of the peptide
in a suitable liquid pharmaceutical carrier or any other formulation such
as tablets, pills, film tablets, coated tablets, dragees, capsules,
powders and deposits, gels, syrups, slurries, suspensions, emulsions, and
the like.

[0201]A particularly preferred pharmaceutical composition is a lyophilised
(freeze-dried) preparation (lyophilisate) suitable for administration by
inhalation or for intravenous administration. To prepare the preferred
lyophilised preparation the peptide of the invention are solubilised in a
4 to 5% (w/v) mannitol solution and the solution is then lyophilised. The
mannitol solution can also be prepared in a suitable buffer solution as
described above.

[0202]Further examples of suitable cryo-/lyoprotectants (otherwise
referred to as bulking agents or stabilizers) include thiol-free albumin,
immunoglobulins, polyalkyleneoxides (e.g. PEG, polypropylene glycols),
trehalose, glucose, sucrose, sorbitol, dextran, maltose, raffinose,
stachyose and other saccharides (cf. for instance WO 97/29782), while
mannitol is used preferably. These can be used in conventional amounts in
conventional lyophilization techniques. Methods of lyophilisation are
well known in the art of preparing pharmaceutical formulations.

[0203]For administration by inhalation the particle diameter of the
lyophilised preparation is preferably between 2 to 5 μm, more
preferably between 3 to 4 μm. The lyophilised preparation is
particularly suitable for administration using an inhalator, for example
the OPTINEB® or VENTA-NEB® inhalator (NEBU-TEC, Elsenfeld,
Germany). The lyophilised product can be rehydrated in sterile distilled
water or any other suitable liquid for inhalation administration.

[0204]Alternatively for intravenous administration the lyophilised product
can be rehydrated in sterile distilled water or any other suitable liquid
for intravenous administration.

[0205]After rehydration for administration in sterile distilled water or
another suitable liquid the lyophilised preparation should have the
approximate physiological osmolality of the target tissue for the
rehydrated peptide preparation i.e. blood for intravenous administration
or lung tissue for inhalation administration. Thus it is preferred that
the rehydrated formulation is substantially isotonic.

[0206]The preferred dosage concentration for either intravenous, oral, or
inhalation administration is between 100 to 2000 μmole/ml, and more
preferably is between 200 to 800 μmole/ml. These are also the
preferred ranges of the peptide in the mother milk substitute or
artificial mother milk formulation or the pharmaceutical compositions
disclosed herein.

Dietary Supplement

[0207]Still another aspect of the present invention relates to the use of
disclosed peptide as a dietary supplement. That dietary supplement is
preferably for oral administration and especially but not limited to
administration to newborns, toddlers, and/or infants.

[0208]A dietary supplement is intended to supplement the diet. The
"dietary ingredients" in these products may in addition include:
vitamins, minerals, herbs or other botanicals, amino acids, and
substances such as enzymes, organ tissues, glandulars, and metabolites.
Dietary supplements may be manufactured in forms such as tablets,
capsules, softgels, gelcaps, liquids, or powders.

Method of Treatment

[0209]Another aspect of the present invention relates to a method of
prophylaxis and/or treatment of cancer, an autoimmune disease, a fibrotic
disease, an inflammatory disease, a neurodegenerative disease, an
infectious disease, a lung disease, a heart and vascular disease or a
metabolic disease or any other disease disclosed herein comprising
administering to a patient in need thereof a pharmaceutical composition
comprising the peptide
Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-OH in a
therapeutically effective amount effective to treat the afore-mentioned
disease.

[0210]Accordingly, the terms "prophylaxis" or "treatment" includes the
administration of the peptide of the present invention to prevent,
inhibit, or arrest the symptoms of an infectious disease, an autoimmune
disease, a fibrotic disease, an inflammatory disease, a neurodegenerative
disease, or a heart and vascular disease. In some instances, treatment
with the peptide of the present invention will be done in combination
with other protective compounds to prevent, inhibit, or arrest the
symptoms of an infectious disease, an autoimmune disease, a fibrotic
disease, an inflammatory disease, a neurodegenerative disease, or a heart
and vascular disease.

[0211]The term "active agent" or "therapeutic agent" as used herein refers
to an agent that can prevent, inhibit, or arrest the symptoms and/or
progression of an infectious, an autoimmune disease, a fibrotic disease,
an inflammatory disease, a neurodegenerative disease, or a heart and
vascular disease or any other disease disclosed herein.

[0212]The term "therapeutic effect" as used herein, refers to the
effective provision of protection effects to prevent, inhibit, or arrest
the symptoms and/or progression of an infectious, an autoimmune disease,
a fibrotic disease, an inflammatory disease, a neurodegenerative disease,
or a heart and vascular disease.

[0213]The term "a therapeutically effective amount" as used herein means a
sufficient amount of the peptide of the invention to produce a
therapeutic effect, as defined above, in a subject or patient in need of
treatment.

[0214]The terms "subject" or "patient" are used herein mean any mammal,
including but not limited to human beings, including a human patient or
subject to which the compositions of the invention can be administered.
The term mammals include human patients and non-human primates, as well
as experimental animals such as rabbits, rats, and mice, and other
animals.

[0215]The peptide of the present invention can be used for the prophylaxis
and/or treatment of cancer, an autoimmune disease, a fibrotic disease, an
inflammatory disease, a neurodegenerative disease, an infectious disease,
a lung disease, a heart and vascular disease or a metabolic disease or
any other disease mentioned herein in combination administration with
another therapeutic compound. As used herein the term "combination
administration" of a compound, therapeutic agent or known drug with the
peptide of the present invention means administration of the drug and the
peptide at such time that both the known drug and the peptide will have a
therapeutic effect. In some cases this therapeutic effect will be
synergistic. Such concomitant administration can involve concurrent (i.e.
at the same time), prior, or subsequent administration of the drug with
respect to the administration of the peptide of the present invention. A
person of ordinary skill in the art would have no difficulty determining
the appropriate timing, sequence and dosages of administration for
particular drugs and peptide of the present invention.

Definition of Peptide Activity

[0216]A peptide is deemed to have therapeutic activity if it demonstrated
any one of the following activities listed in a) to g).

a) The peptide could inhibit the activity of an over active biological
pathway.b) The peptide could inhibit the production of an over produced
biological molecule.c) The peptide could inhibit the activity of an over
produced biological molecule.d) The peptide could increase the activity
of an under active biological pathway.e) The peptide could increase the
production of an under produced biological molecule.f) The peptide could
mimic the activity of an under produced biological molecule.g) The
peptide could prevent, inhibit, or arrest the symptoms and/or progression
of cancer, an infectious disease, an autoimmune disease, a fibrotic
disease, an inflammatory disease, a neurodegenerative disease, or a heart
and vascular disease or any other disease disclosed herein.

[0217]As used herein "inhibition" is defined as a reduction of the
activity or production of a biological pathway or molecule activity of
between 10 to 100%. More preferably the reduction of the activity or
production of a biological pathway or molecule activity is between 25 to
100%. Even more preferably the reduction of the activity or production of
a biological pathway or molecule activity is between 50 to 100%.

[0218]As used herein "increase" is defined as an increase of the activity
or production of a biological pathway or molecule of between 10 to 100%.
More preferably the increase of the activity or production of a
biological pathway or molecule activity is between 25 to 100%. Even more
preferably the increase of the activity or production of a biological
pathway or molecule activity is between 50 to 100%.

[0219]As used herein "mimic" is defined as an increase in the activity of
a biological pathway dependent on the under produced biological molecule
of between 10 to 100%. More preferably the increase of the activity of
the biological pathway is between 25 to 100%. Even more preferably the
increase of the activity the biological pathway is between 50 to 100%.

Peptide

[0220]The peptide of the invention was for tested for the activity as a
therapeutic agent for the prophylaxis and/or treatment of cancer, an
infectious disease, an autoimmune disease, a fibrotic disease, an
inflammatory disease, a neurodegenerative disease, or a heart and
vascular disease:

[0221]The term "TRAP-14" in brackets after the peptide sequence
Ser-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-OH is an
abbreviation or synonym of said peptide.

[0222]Furthermore the present invention relates to the use of the
above-mentioned peptide as pharmaceutically active agents in medicine,
i.e. as medicament. Advantage of the peptide of the invention is that the
peptide is less toxic in comparison to the commonly used drugs for the
certain indications mentioned herein and that the peptide have less side
effects, can be used for a long term treatment of certain diseases and
can be easily administered. Moreover the peptide are selective for
certain targets and under physiological conditions no toxic or noxious
degradation products are formed.

[0223]As used herein, the term "peptide(s)" or "peptide(s) of the
invention" shall also refer to salts, deprotected form, acetylated form
of the peptide, deacetylated form of the peptide, enantiomers,
diastereomers, racemates, prodrugs and hydrates of the above-mentioned
peptide. Diastereomers of the peptide are obtained when the
stereochemical or chiral center of one or more amino acids is changed.
The enantiomer has the opposite stereochemistry at all chiral centers.

[0224]The term "prodrug" refers to any precursor compound which is able to
generate or to release the above-mentioned peptide under physiological
conditions. Such prodrugs, i.e. such precursor molecules are for instance
larger peptides which are selectively cleaved in order to form the
peptide of the invention. Further prodrugs are protected amino acids
having especially protecting groups at the carboxylic acid and/or amino
group.

[0225]Suitable protecting groups for amino groups are the
benzyloxycarbonyl, t-butyloxycarbonyl (BOC), formyl, and acetyl or acyl
group. Suitable protecting groups for the carboxylic acid group are
esters such as benzyl esters or t-butyl esters.

[0226]The present invention also includes the above peptide having amino
acid substitutions, deletions, additions, the substitutions and additions
including the standard D and L amino acids and modified amino acids such
as for example amidated and acetylated amino acids, wherein the
therapeutic activity of the base peptide sequence as shown above is
maintained.

"D-2-NaI" is 2-naphthyl-D-alanine,"SertBu" is t-butyl serine,"Azagly" is
aza glycine,"Me" is methyl,Met(O) is methionine sulfoxide,"Pyr" and
"pGlu" are pyroglutamic acid,"Tyr(SO3H)" is sulphated
tyrosine,"Tyr(Me)" is methyltyrosine,"NHEt" is ethylamide.

EXAMPLES

[0229]The peptides as listed above were tested for activity using the
assays described in Examples 1 to 17. The tested peptides are all
commercially available.

Example 1

HIV-1 Experiments

[0230]CEM-SS cells were passaged in T-75 flasks prior to use in the
antiviral assay. On the day preceding the assay, the cells were split 1:2
to assure they were in an exponential growth phase at the time of
infection. Total cell viability quantification was performed using a
hemacytometer and trypan blue exclusion. Cell viability was greater than
95% for the cells to be utilized in the assay. The cells were resuspended
at 5×104 cells/ml in tissue culture medium and added to the
peptide-containing microtiter plates in a volume of 50 microliters.

[0231]The virus used was the lymphocytotropic strain HIV-1IIIB. Virus
was obtained from NIH AIDS Research and Reference Reagent Program and was
grown in CEM-SS cells for the production of stock virus pools. For each
assay, a pre-titered aliquot of virus was removed from the freezer
(-80° C.) and allowed to thaw slowly to room temperature in a
biological safety cabinet. The virus was resuspended and diluted into
tissue culture medium such that the amount of virus added to each well in
a volume of 50 microliters was the amount determined to give between 85%
to 95% cell killing after 6 days post-infection. TCID50 calculations
by endpoint titration in CEM-SS cells indicated that the multiplicity of
infection was approximately 0.01. AZT (nucleoside reverse transcriptase
inhibitor; NRTI) and indinavir (protease inhibitor; PI) were used as
positive control antiviral compounds.

[0233]At assay termination, the plates were stained with the soluble
tetrazolium-based dye MTS (CellTiter 96 Reagent, Promega) to determine
cell viability and quantify peptide toxicity. MTS is metabolized by the
mitochondrial enzymes of metabolically active cells to yield a soluble
formazan product, allowing the rapid quantitative analysis of cell
viability and peptide cytotoxicity. This reagent is a stable, single
solution that does not require preparation before use. At assay
termination, 20-25 microliters of MTS reagent was added per well and the
microtiter plates were then incubated for 5 hours at 37° C., and
5% CO2 to assess cell viability. Adhesive plate sealers were used in
place of lids, the sealed plates were inverted several times to mix the
soluble formazan product and the plate was read spectrophotometrically at
490/560 nm with a Molecular Devices Vmax plate reader.

[0234]The overall assay performance was valid based upon judgement of the
positive control compounds AZT and indinavir exhibiting the expected
levels of antiviral activity. Macroscopic observation of the cells in
each well of the microtiter plate confirmed the cytotoxicity results
obtained following staining of the cells with the MTS metabolic dye.

[0236]HepG2-2.2.15 is a stable cell line containing the hepatitis B virus
(HBV) ayw strain genome (ATCC Cat. No. CRL-11997). Antiviral compounds
blocking any late step of viral replication such as transcription,
translation, pregenome encapsidation, reverse transcription, particle
assembly and release can be identified and characterized using this cell
line. In this assay, an active compound will reduce the production of
secreted HBV from cells, measured by utilizing real time quantitative PCR
(TaqMan) assay to directly and accurately measure HBV DNA copies. The
analysis of this data allows to calculate: [0237]Antiviral activity
[0238]Compound Cytotoxicity

[0239]HepG2-2.2.15 cells were plated in 96-well microtiter plates. After
16-24 hours the confluent monolayer of HepG2-2.2.15 cells was washed and
the medium was replaced with complete medium containing test peptide--10
micrograms per ml--in duplicate. Lamivudine (3TC) was used as the
positive control, while media alone was added to the cells as a negative
control (virus control). Three days later the culture medium was replaced
with fresh medium containing the peptide. Six days following the initial
administration of the peptide, the cell culture supernatants was
collected, treated with pronase and DNAse and then used in a real-time
quantitative TaqMan PCR assay. The PCR-amplified HBV DNA was detected in
real-time by monitoring increases in fluorescence signals that result
from the exonucleolytic degradation of a quenched fluorescence probe
molecule that hybridizes to the amplified HBV DNA. For each PCR
amplification, a standard curve was simultaneously generated using
dilutions of purified HBV DNA. Antiviral activity was calculated from the
reduction in HBV DNA levels (% virus control). A novel dye uptake assay
was then employed to measure cell viability, which is used to calculate
toxicity (% cell control).

[0242]MRC-5 cells were seeded at 75,000 cells/well in 24 well plates using
MRC-5 growth medium. The plates were incubated overnight at 37°
C., 5% CO2. The following day, media was removed and 100 plaque
forming units (pfu) of HCMV was added to the wells. Virus was allowed to
adsorb onto the cells for 1 hour at 37° C., 5% CO2. Peptide
was diluted--10 micrograms per ml--in assay medium containing 0.5%
Methylcellulose. After the incubation period, 1 ml of each peptide
solution was added to the wells without aspirating the virus inoculums.
The plates were incubated for 7-10 days to allow for plaque formation.
Ganciclovir was used as positive control. Cultures were examined
microscopically and toxicities were noted. The media was the aspirated
from the wells and the cells were fixed and stained using 20% methanol
containing Crystal Violet followed by enumeration of plaques by
microscopic inspection.

[0243]For cytotxicity testing, MRC-5 cells were seeded at 2,500 cells/well
in 96 well plates using growth medium. The plates were incubated
overnight at 37° C., 5% CO2. The following day, peptide was
added and tested in duplicates. After a 6 days incubation period, cell
viability was measured using CellTiter 96 Solution (Promega). Plates were
incubated for additional 4 hours at 37° C. Adhesive plate sealers
were used in place of lids, the sealed plates were inverted several times
to mix the soluble formazan product and the plate was read
spectrophotometrically at 490/560 nm with a Molecular Devices Vmax plate
reader.

[0244]The overall assay performance was valid based upon judgement of the
positive control compound Ganciclovir exhibiting the expected levels of
antiviral activity. Macroscopic observation of the cells in each well of
the microtiter plate confirmed the cytotoxicity results obtained
following staining of the cells with the MTS metabolic dye.

[0246]The antibacterial assay was conducted using clear, U-bottom 96-well
microtiter plates. Cation-adjusted Mueller-Hinton Broth (MHB) was used
for testing MRSA. The peptide of the invention (0.1 ml of each--10
micrograms per ml--) was dispensed into wells in duplicate. Then the
wells were inoculated with 5×105 CFU/mL MRSA in 0.1 ml volume.
For control purposes, each plate included 4 wells containing media
without bacterial inoculum and 4 wells containing medium with inoculum
but without peptide. The plates were incubated for 12 h at 37° C.,
and read visually 18-24 hours post-incubation. Growth control of MRSA was
examined first to determine adequacy of media preparations and growth
conditions. Acceptable growth is defined as 2 mm wide button of cells at
the bottom of each sample well, or obvious turbidity in the culture
supernatant. Test wells were examined and scored as positive/negative for
activity. A positive score for activity is based on complete inhibition
of macroscopic growth of the test MRSA.

Results from MRSA assay:

TABLE-US-00008
Compound % inhibition
TRAP-14 0.0

Example 5

Pseudomonas aeruginosa Assay

[0247]The antibacterial assay was conducted using clear, U-bottom 96-well
microtiter plates. Cation-adjusted Mueller-Hinton Broth (MHB) was used
for testing Pseudomonas aeruginosa. The peptide of the invention (0.1 ml
of each--10 micrograms per ml--) was dispensed into wells in duplicate.
Then the wells were inoculated with 5×105 CFU/mL Pseudomonas
aeruginosa in 0.1 ml volume. For control purposes, each plate included 4
wells containing media without bacterial inoculum and 4 wells containing
medium with inoculum but without peptide. The plates were incubated for
12 h at 37° C., and read visually 18-24 hours post-incubation.
Growth control of Pseudomonas aeruginosa was examined first to determine
adequacy of media preparations and growth conditions. Acceptable growth
is defined as 2 mm wide button of cells at the bottom of each sample
well, or obvious turbidity in the culture supernatant. Test wells were
examined and scored as positive/negative for activity. A positive score
for activity is based on complete inhibition of macroscopic growth of the
test Pseudomonas aeruginosa.

Results from Pseudomonas aeruginosa assay:

TABLE-US-00009
Compound % inhibition
TRAP-14 0.0

Example 6

Streptococcus pneumoniae Assay

[0248]The antibacterial assay was conducted using clear, U-bottom 96-well
microtiter plates. Cation-adjusted Mueller-Hinton Broth (MHB) was used
for testing Streptococcus pneumoniae. The peptide of the invention (0.1
ml of each--10 micrograms per ml--) was dispensed into wells in
duplicate. Then the wells were inoculated with 5×105 CFU/mL
Streptococcus pneumoniae in 0.1 ml volume. For control purposes, each
plate included 4 wells containing media without bacterial inoculum and 4
wells containing medium with inoculum but without peptide. The plates
were incubated for 12 h at 37° C., and read visually 18-24 hours
post-incubation. Growth control of Streptococcus pneumoniae was examined
first to determine adequacy of media preparations and growth conditions.
Acceptable growth is defined as ≧2 mm wide button of cells at the
bottom of each sample well, or obvious turbidity in the culture
supernatant. Test wells were examined and scored as positive/negative for
activity. A positive score for activity is based on complete inhibition
of macroscopic growth of the test Streptococcus pneumoniae.

Results from Streptococcus pneumoniae assay:

TABLE-US-00010
Compound % inhibition
TRAP-14 0.0

Example 7

Mycobacterium tuberculosis Assay

[0249]The antibacterial assay was conducted using clear, U-bottom 96-well
microtiter plates. Middlebrook 7H12 assay medium was used for testing
drug-resistant Mycobacterium tuberculosis. The peptide of the invention
(0.1 ml of each--10 micrograms per ml--) was dispensed into wells in
duplicate. Then the wells were inoculated with 5×105 CFU/mL
Mycobacterium tuberculosis in 0.1 ml volume. For control purposes, each
plate included 4 wells containing media without bacterial inoculum and 4
wells containing medium with inoculum but without peptide. The plates
were incubated for seven days at 37° C., and read visually
thereafter. Growth control of Mycobacterium tuberculosis was examined
first to determine adequacy of media preparations and growth conditions.
Acceptable growth is defined as 2 mm wide button of cells at the bottom
of each sample well, or obvious turbidity in the culture supernatant.
Test wells were examined and scored as positive/negative for activity. A
positive score for activity is based on complete inhibition of
macroscopic growth of the test Mycobacterium tuberculosis. The
drug-resistant Mycobacterium tuberculosis that was used in the assay is
resistant against following medicaments: para-aminosalicylic acid (PAS),
streptomycin and isoniazid (INH).

Results from Mycobacterium tuberculosis assay:

TABLE-US-00011
Compound % inhibition
TRAP-14 0.0

Example 8

Cell Cycle Assay

[0250]Human A549 cells (carcinomic human alveolar basal epithelial cells)
were utilized in the experiments employing the Propidium iodide cell
cycle assay. The eukaryotic cell cycle is a series of events that take
place in a cell leading to its replication.

[0251]The regulation of the cell cycle involves steps crucial to the cell,
including detecting and repairing genetic damage, and provision of
various checks to prevent uncontrolled cell division. The molecular
events that control the cell cycle are ordered and directional; that is,
each process occurs in a sequential fashion.

[0252]The cell cycle consists of four distinct phases: G1 phase, S
phase, G2 phase (collectively known as interphase) and M phase. M
phase is itself composed of two tightly coupled processes: mitosis, in
which the cell's chromosomes are divided between the two daughter cells,
and cytokinesis, in which the cell's cytoplasm divides forming distinct
cells. Activation of each phase is dependent on the proper progression
and completion of the previous one. Cells that have temporarily or
reversibly stopped dividing are said to have entered a state of
quiescence called Go phase. The relatively brief M phase consists of
nuclear division and cytoplasmic division. The first phase within
interphase, from the end of the previous M phase till the beginning of
DNA synthesis is called G1 (G indicating gap or growth). During this
phase the biosynthetic activities of the cell resume at a high rate. This
phase is marked by synthesis of various enzymes that are required in S
phase, mainly those needed for DNA replication. The ensuing S phase
starts when DNA synthesis commences; when it is complete, all of the
chromosomes have been replicated. The cell then enters the G2 phase,
which lasts until the cell enters mitosis. Significant protein synthesis
occurs during this phase, mainly involving the production of
microtubules, which are required during the process of mitosis.
Inhibition of protein synthesis during G2 phase prevents the cell
from undergoing mitosis. Disregulation of the cell cycle components may
lead to tumor formation.

[0253]Propidium iodide is an intercalating agent and a fluorescent
molecule that can be used to stain DNA. Cells were incubated for 24 hours
with test peptide--10 micrograms per ml--or left untreated. After that
cells were trypsinized, suspended in medium+10% FCS, Centrifuged (1000
rpm, 5 min), and the cell pellet resuspended in PBS (1 ml). The cells
were pipetted into 2.5 ml absolute EtOH (final concentration approx. 70%)
and incubated on ice for 15 min. Thereafter, cells were pelleted at 1500
rpm for 5 min and resuspended in Propidium iodide solution in PBS. After
incubation for 40 min at 37° C., cells were analyzed in the FACS.

[0254]Human Peripheral Blood Mononuclear Cells (PBMC) were obtained from
normal human donors. The T cell proliferation was induced by stimulation
of the cells with the T cell mitogen phytohemagglutinin (PHA), either in
the absence (positive proliferation control), or in the presence of test
peptide--10 micrograms per ml--to examine their effects on the T cell
proliferating response. 105/well PBMC were plated in 96-well
microtiter plates and assayed in duplicate with the peptide. Cell
cultures were incubated at 37° C. for 3 days in a 5% CO2
incubator and were thereafter pulsed with 1 microCi/well
3H-thymidine for additional 12 hours of culture. At the end of
incubation time, the plates were harvested and the cells counted by
liquid scintillation for the incorporation of 3H-thymidine as a
measure of T cell proliferation.

Results from T cell proliferation assay:

TABLE-US-00013
% of PHA induced
Compound Control
TRAP-14 95.7

Example 10

B Cell Proliferation Assay

[0255]Human Peripheral Blood Mononuclear Cells (PBMC) were obtained from
normal human donors. The B cell proliferation was induced by stimulation
of the cells with the B cell mitogen Staphylococcus aureus Cowans I (SAC)
plus Interleukin-2, either in the absence (positive proliferation
control), or in the presence of test peptide--10 micrograms per ml--to
examine their effects on the B cell response. 105/well PBMC were
plated in 96-well microtiter plates and assayed in duplicate with the
peptide. Cell cultures were incubated at 37° C. for 3 days in a 5%
CO2 incubator and were thereafter pulsed with 1 microCi/well
3H-thymidine for additional 12 hours of culture. At the end of
incubation time, the plates were harvested and the cells counted by
liquid scintillation for the incorporation of 3H-thymidine as a
measure of B cell proliferation.

Results from B cell proliferation assay:

TABLE-US-00014
% of SAC/IL2
induced
Compound control
TRAP-14 108.8

Example 11

Phagocytosis Assay

[0256]RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) cells were
obtained from ATCC and grown in RPMI 1640 medium containing 10% FBS.
Cells were incubated in 12×75 mm tubes at 37° C. with test
peptide--10 micrograms per ml--for 30 min prior to adding
Fluorescein-labeled Escherichia coli bacteria as the agent to be
ingested. After the cells were incubated for additional 60 min at
37° C. and allowed to ingest the Fluorescein-labeled Escherichia
coli bacteria, cells were fixed with 1% paraformaldehyde. The samples
were then analyzed by flow cytometry to determine the amount of
phagocytosis as a function of brightness (the greater the phagocytic
activity, the more fluorescence in the macrophage population). Data are
reported as % positive and the mean fluorescence intensity (MFI) of
positively stained cells.

Results from phagocytosis assay:

TABLE-US-00015
% of control
Compound phagocytosis
TRAP-14 105.9

Example 12

Apoptosis Induction Assay

[0257]Human A549 cells (carcinomic human alveolar basal epithelial cells)
were utilized in the experiments employing the Annexin-5 apoptosis assay.
Annexin-5 is a member of a highly conserved protein family that binds
acidic phospholipids in a calcium-dependent manner. Annexin-5 possesses a
high affinity for phosphatidylserine. Phosphatidylserine is translocated
from the inner side of the plasma membrane to the outer layer when cells
undergo death by apoptosis or cell necrosis and serves as a signal by
which cell destined for death are recognized by phagocytes. Test
peptide--10 micrograms per ml--were exposed for 24 hours to the A549
cells before they were analyzed for signs of apoptosis.

Results from apoptosis induction assay:

TABLE-US-00016
Compound % of induction
TRAP-14 0.3

Example 13

Apoptosis Prevention Assay

[0258]Human A549 cells (carcinomic human alveolar basal epithelial cells)
were utilized in the experiments employing the Annexin-5 apoptosis assay.
Annexin-5 is a member of a highly conserved protein family that binds
acidic phospholipids in a calcium-dependent manner. Annexin-5 possesses a
high affinity for phosphatidylserine. Phosphatidylserine is translocated
from the inner side of the plasma membrane to the outer layer when cells
undergo death by apoptosis or cell necrosis and serves as a signal by
which cell destined for death are recognized by phagocytes. A549 cells
were pretreated for 30 min with test peptide--10 micrograms per
ml--followed by the exposure to C2 ceramide. Ceramide mediates cell
apoptosis through the activation of the mitogen activating protein kinase
(MAPK) and the stress activated kinase (JNK/SAPK). C2 ceramide is a
synthetic, membrane soluble analog of ceramide.

[0259]The Balb/c mice (originated in 1923, it is a popular strain and is
used in many different research disciplines. Also classified as an inbred
from the production of 20 or more successive brother-sister matings, the
Balb/c mouse is albino and small in size) were immunized on Days 1, 15,
and 29 with Ovalbumin (Ovalbumin is the main protein found in egg white,
commonly used to stimulate an immunological reaction in test animals) in
PBS (5 micrograms/injection). On day 50, spleens of the mice were
harvested (3 weeks after last boost with Ovalbumin). Cells were cultured
(2×105/well in triplicate) and incubated with culture medium
or test peptide--10 micrograms per ml--for 30 min. Thereafter, additional
Ovalbumin was added to the cells at 10 micrograms/ml for in vitro
restimulation of the cells. 72 hours later, cell supernatants were
harvested and assayed using the Becton Dickinson Mouse Th1/Th2 Cytokine
CBA Kit. This kit can be used to measure Interleukin-2 (IL-2),
Interleukin-4 (IL-4), Interleukin-5 (IL-5), Interferon-γ
(IFN-γ), and Tumor Necrosis Factor-α (TNF-α) protein
levels in a single sample. The kit performance has been optimized for
analysis of physiologically relevant concentrations (pg/ml levels) of
specific cytokine proteins in tissue culture supernatants and serum
samples.

[0260]Human Peripheral Blood Mononuclear Cells (PBMC) were obtained from
normal human donors. The macrophages were prepared by adherence of PBMC
to the plastic wells of the plates. After 8 days in culture in the
presence of recombinant human macrophage-colony stimulating factor at 2
ng/ml, differentiated macrophages were preincubated with test peptide--10
micrograms per ml--for 30 min, followed by in-well stimulation by the
addition of lipopolysaccharide at a final concentration of 200 ng/ml. Not
stimulated macrophages served as negative background control. After
overnight incubation, supernatants from the control and LPS-stimulated
cultures were harvested and assayed for TNF alpha production employing a
TNF alpha specific ELISA.

Results from TNF alpha assay:

TABLE-US-00019
Compound % of LPS induction
TRAP-14 110.0

Example 16

Endothelial Cell Migration Assay

[0261]Endothelial cell migration is a prerequisite for the process of
neo-vascularization or angiogenesis which is crucial for on-site
recruitment of blood vessel formation. Primary Human endothelial cells
(HUVEC) were seeded in insert chambers with 3 micrometer pore size of
multi-transwell plate for 6 hours at 37° C. in Endothelial Cell
Basal Medium (EBM) supplemented with 0.1% bovine serum albumin.
Thereafter, designated concentration of test peptide--10 micrograms per
ml--was added in duplicate wells. The endothelia were allowed to migrate
for 22 hours at 37° C., then, migrated cells were fixed and
stained with Hoechst 33342 dye. Images of 3 fields per insert were taken
and the number of migrated cells per field were quantified using the
ImageProPlus software. Data were analyzed for the average number of the
migrated cells and standard deviation of six data points for each
treatment condition. Active test peptide against HUVEC migration was
determined based on 50% inhibition of migrated cells as compared with the
control. Statistic p values were computed using the Student's t-test.

Results from endothelial cell migration assay:

TABLE-US-00020
% inhibition of
Compound migration
TRAP-14 -45

Example 17

Endothelial Tube Formation Assay

[0262]The endothelial tube formation assay is based on the ability of
endothelial cells to form three-dimensional capillary-like tubular
structures when cultured on a gel of basement membrane extract. The
endothelial tube formation assay represents a powerful model for studying
inhibition and induction of angiogenesis. Pre-labeled HUVEC with Calcein
AM were seeded in a 96-well culture plate coated with extracellular
metrix (Chemicon international Cat. ECM625) and treated with test
peptide--10 micrograms per ml--in full growth medium. Positive control
was vehicle only. The endothelial cells were allowed to form tubes for 20
hours and were then examined under an inverted fluorescent microscope.
Duplicate wells for each treatment were photographed and quantitatevily
analyzed for an average tubule length using image analysis software
ImageProPlus. Raw data were expressed as average tubule lengths in
pixels±standard deviation. Statistic p values were computed using the
Student's t-test.

Results from endothelial tube formation assay:

TABLE-US-00021
% inhibition of tube
Compound formation
TRAP-14 16

Example 18

cAMP Induction Assay

[0263]The cAMP kit was used as an immunoassay for the quantitative
determination of cyclic AMP generation in human peripheral blood
mononuclear cells (PBMC). PBMC were placed in 96-well microtitier plates
(50 microliters of cells at 4×106/ml) and 50 microliters of
medium alone (background control), forskolin (cAMP positive control; 50
micrograms per ml), or test peptide were added. Plates were mixed and
incubated for 10 min at 37° C. and the cells were lysed by the
addition of 50 microliters of 3% perchloric acid. NaOH was added to
neutralize the samples. Plates were centrifuged for 15 min at 2500 rpm
and the supernatants removed. The supernatants (diluted 2-fold in assay
buffer) were then assayed for the levels of cAMP by ELISA.

[0265]One example of an artificial mother milk or mother milk substitute
formulation is provided in the following while also the other
formulations disclosed in the above mentioned references can be used and
are included herewith by reference.

[0267]In a first step, the solubilizing aqueous medium is produced,
comprises, by weight, approximately 75% of water, approximately 0.02% of
carrageenate and approximately 0.2% of disodium hydrogenphosphate.

[0268]The skimmed milk powder is then added to the solution for 10 min at
60° C. and dissolved in the liquid.

[0269]Then soya oil and lecithin are added to the milk substitute
composition at 60° C. The milk composition is allowed to stand 30
min at 55° C. After pasteurization, the peptide of the invention
is added in liquid or powder form in such a quantity that the milk
composition obtained comprises an amount of 5-50 micrograms, preferably
10-40 micrograms per 100 ml of milk composition. Optionally peptide 2
could be added in similar or smaller amounts to the obtained composition.

Example 20

Gel Formulation

[0270]0.5 g of peptide1.6 g of isopropanol1.0 g of glycerol1.6 g of
polyoxyethylene-polyoxypropylene copolymer 12500 (Pluronic F127)5.3 g of
waterare mixed for 10 minutes and then heated to 85° C. under
continuous stirring for 15 minutes. The solution is cooled to room
temperature under stirring. During the cooling phase the solution begins
to gel at a temperature of about 45° C. to form a clear gel. The
gel contains 5% of the peptide 1 for medical use. Optionallly peptide 2
could be added in an amount form 0.01 to 0.5 g.

Example 21

Lotion Formulation

[0271]0.5 g of peptide1.9 g of isopropanol1.0 g of dimethylisosorbide1.0 g
of polyoxyethylene-polyoxypropylene copolymer 12500 (Pluronic F127)5.6 g
of waterare stirred and heated at 50° C., until a clear solution
has been formed. Then the composition is cooled to room temperature under
stirring. The lotion contains 5% of peptide 1 for medical use. Optionally
peptide 2 could be added in an amount form 0.01 to 0.5 g.